Transcriptome of GH-producing pituitary neuroendocrine tumours and models are significantly affected by somatostatin analogues

Pituitary neuroendocrine tumours (PitNETs) are neoplasms of the pituitary that overproduce hormones or cause unspecific symptoms due to mass effect. Growth hormone overproducing GH-producing PitNETs cause acromegaly leading to connective tissue, metabolic or oncologic disorders. The medical treatment of acromegaly is somatostatin analogues (SSA) in specific cases combined with dopamine agonists (DA), but almost half of patients display partial or full SSA resistance and potential causes of this are unknown. In this study we investigated transcriptomic landscape of GH-producing PitNETs on several levels and functional models—tumour tissue of patients with and without SSA preoperative treatment, tumour derived pituispheres and GH3 cell line incubated with SSA to study effect of medication on gene expression. MGI sequencing platform was used to sequence total RNA from PitNET tissue, pituispheres, mesenchymal stromal stem-like cells (MSC), and GH3 cell cultures, and data were analysed with Salmon—DeSeq2 pipeline. We observed that the GH-producing PitNETs have distinct changes in growth hormone related pathways related to its functional status alongside inner cell signalling, ion transport, cell adhesion and extracellular matrix characteristic patterns. In pituispheres model, treatment regimens (octreotide and cabergoline) affect specific cell proliferation (MKI67) and core functionality pathways (RYR2, COL8A2, HLA-G, ARFGAP1, TGFBR2). In GH3 cells we observed that medication did not have transcriptomic effects similar to preoperative treatment in PitNET tissue or pituisphere model. This study highlights the importance of correct model system selection for cell transcriptomic profiling and data interpretation that could be achieved in future by incorporating NGS methods and detailed cell omics profiling in PitNET model research. Supplementary Information The online version contains supplementary material available at 10.1186/s12935-023-02863-4.

Summarization of the categories in enrichment analysis results for the PitNET non-secreting and somatotroph type comparison. Each bar represents an enrichment category with the corresponding occurance in enrichment results.

Figure S2.
A STRING-db network for the DEG proteins of somatotroph and nonsecreting PitNET type comparison. Line thickness indicates confidence of protein interaction. Disconnected nodes were hidden. Text mining disabled as interaction type.

Figure S3.
A STRING db network for the DEG proteins of somatotroph and nonsecreting PitNET type comparison. Line colour indicates protein interaction type and line count indicates number of interactions. Disconnected nodes were hidden. Text mining disabled as interaction type.

Figure S4.
A STRING -db network for the DEG proteins of SSA treated/untreted somatotroph PitNET comparison. Line thickness indicates interaction confidence. Disconnected nodes were hidden. Text mining disabled as interaction type.  Variance stabilization transformed heatmap of mean normalized gene level counts for DEGs of the octreotide incubated pituispheres cell and pituisphere control comparsion at 4 hours. Bright green (middle) represents samples from the octreotide 4h incubation period, darker green represents the unincubated pituisphere control samples.

Figure S7.
Variance stabilization transformed heatmap of mean normalized gene level counts for DEGs of the octreotide incubated pituispheres cell and pituisphere control comparsion at 8 hours. Dark green represents the unincubated pituisphere control samples, bright green represents the octreotide incubated pituispheres at 8 hours.

Figure S8.
Variance stabilization transformed heatmap of mean normalized gene level counts for DEGs of the octreotide incubated pituispheres cell and pituisphere control comparsion at 24 hours. Dark green represents the unincubated pituisphere control samples, bright green represents the octreotide incubated pituispheres at 24 hours.

Figure S9.
Variance stabilization transformed heatmap of mean normalized gene level counts for DEGs of the octreotide incubated pituispheres cell and pituisphere control comparsion at 48 hours. Dark green represents the unincubated pituisphere control samples, bright green represents the octreotide incubated pituispheres at 48 hours.       Variance stabilization transformed heatmap of mean normalized gene level counts for DEGs of the octreotide incubated GH3 cell and GH3 control comparsion at 4 hours. Light blue fields in the "Time" column row, represent the unincubated control samples, orange fields represent the GH3 samples incubated with octreotide at 4 hours.

Figure S17.
Variance stabilization transformed heatmap of mean normalized gene level counts for DEGs of the octreotide incubated GH3 cell and GH3 control comparsion at 8 hours. Light blue fields in the "Time" column row, represent the unincubated control samples, orange fields represent the GH3 samples incubated with octreotide at 8 hours.

Figure S18.
Variance stabilization transformed heatmap of mean normalized gene level counts for DEGs of the octreotide incubated GH3 cell and GH3 control comparsion at 24 hours. Light blue fields in the "Time" column row, represent the unincubated control samples, orange fields represent the GH3 samples incubated with octreotide at 24 hours.

Figure S19.
A STRING-db network for the DEG proteins of octreotide incubated GH3 cell and GH3 control comparsion for the incubation period of 24 hours. Line thickness indicates interaction confidence. Disconnected nodes were hidden. Text mining disabled as interaction type.

Figure S20.
A STRING-db network for the DEG proteins of octreotide incubated GH3 cell and GH3 control comparsion for the incubation period of 24 hours. Line colour indicated interaction type and line count indicates number of interactions. Disconnected nodes were hidden. Text mining disabled as interaction type. Figure S21.